LOCUS       sequence-396788-        6981 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector for closed-loop ADAR-based RNA sensors (DART VADAR). 
            Expresses TagBFP constitutively. Expresses mNeonGreen and MCP-ADAR 
            upon target detection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6981)
  AUTHORS   Gayet RV, Ilia K, Razavi S, Tippens ND, Lalwani MA, Zhang K, Chen 
            JX, Chen JC, Vargas-Asencio J, Collins JJ
  TITLE     Autocatalytic base editing for RNA-responsive translational control.
  JOURNAL   Nat Commun. 2023 Mar 11;14(1):1339. doi: 10.1038/s41467-023-36851-z.
  PUBMED    36906659
REFERENCE   2  (bases 1 to 6981)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2023-03-11"; volume: "14(1):1339. doi"; pages: " 
            10.1038/s41467-023-36851-z"
COMMENT     Sequence Label: 
            pDV06_feedback_entry_TagBFP-2A-MCS-2A-mNeonGreen-2A-MCP-ADAR2dd(E488
            Q)-NES
FEATURES             Location/Qualifiers
     source          1..6981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        348..559
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          560..1498
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1455..1475
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2246..2302
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2366..2419
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      2681..2690
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3143..3202
                     /codon_start=1
                     /product="2A peptide from equine rhinitis A virus 
                     polyprotein"
                     /label=E2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="QCTNYALLKLAGDVESNPGP"
     CDS             4808..4834
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     polyA_signal    4889..4937
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5214..5802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5294..5313)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5973..6764,6765..6833))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6596..6615
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6834..6938)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgcgttttta tgcttgtagt attgtataat gtttttaaga tccggagaga gcgagattcc
       61 gtctcaaaga aaaaaaaagt aatgaaatga ataaaatgag tcctagagcc agtaaatgtc
      121 gtaaatgtct cagctagtca ggtagtaaaa ggtctcaact aggcagtggc agagcaggat
      181 tcaaattcag ggctgttgtg atgcctccgc agactctgag cgccacctgg tggtaatttg
      241 tctgtgcctc ttctgacgtg gaagaacagc aactaacaca ctaacacggc atttactatg
      301 ggccagccat tgttctaata ctttaattgg ctccggtgcc cgtcagtggg cagagcgcac
      361 atcgcccaca gtccccgaga agttgtgggg aggggtcggc aattgaaccg gtgcctagag
      421 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
      481 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
      541 gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac
      601 gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg tgattcttga
      661 tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt aaggagcccc
      721 ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg tgcgaatctg
      781 gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt aaaatttttg
      841 atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg gccaagatct
      901 gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg
      961 cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac gggggtagtc
     1021 tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg
     1081 ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg
     1141 ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg cgggtgagtc
     1201 acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg actccacgga
     1261 gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta cgtcgtcttt
     1321 aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg tggagactga
     1381 agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg
     1441 atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc catttcaggt
     1501 ggatcccaag ggcagataag tttgtacaaa aaagagaggt gccaccatga gcgagctgat
     1561 taaggagaac atgcacatga agctgtacat ggagggcacc gtggacaacc atcacttcaa
     1621 gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt
     1681 ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctactagct tcctctacgg
     1741 cagcaagacc ttcatcaacc acacccaggg catccccgac ttcttcaagc agtccttccc
     1801 tgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac
     1861 ccaggacacc agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa
     1921 cttcacatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg ccttcaccga
     1981 gacgctgtac cccgctgacg gcggcctgga aggcagaaac gacatggccc tgaagctcgt
     2041 gggcgggagc catctgatcg caaacatcaa gaccacatat agatccaaga aacccgctaa
     2101 gaacctcaag atgcctggcg tctactatgt ggactacaga ctggaaagaa tcaaggaggc
     2161 caacaacgag acatacgtcg agcagcacga ggtggcagtg gccagatact gcgacctccc
     2221 tagcaaactg gggcacggta gcggcgcaac aaacttctct cttctgaaac aggcaggaga
     2281 tgtggaagaa aatccaggcc caggatccgg tttaattaag ggcctgcagg gtggaaagct
     2341 ttctactcga ggggctagcc agggagaggg cagaggaagt ctgcttacat gcggtgatgt
     2401 cgaagagaat cctggcccag gtaccatggt gtccaagggc gaagaggaca acatggccag
     2461 cctgcctgcc acccacgagc tgcacatctt cggcagcatc aacggcgtgg acttcgacat
     2521 ggtgggacag ggcaccggca accccaacga cggatacgag gaactgaacc tgaagtccac
     2581 aaagggcgac ctgcagttca gcccctggat cctggtgccc cacatcggct acggcttcca
     2641 ccagtacctg ccctaccctg acggcatgag ccctttccag gccgccatgg tggatggctc
     2701 tggctaccag gtgcacagaa ccatgcagtt cgaggacggc gccagcctga ccgtgaacta
     2761 cagatacacc tacgagggca gccacatcaa gggcgaggcc caagtgaagg gcacaggctt
     2821 ccctgctgac ggccccgtga tgaccaactc tctgacagcc gccgactggt gcagaagcaa
     2881 gaaaacctac cctaacgaca agaccatcat cagcaccttc aagtggtcct acaccacagg
     2941 caacggcaag agatacagaa gcaccgccag aaccacctac accttcgcca agcccatggc
     3001 cgccaactac ctgaagaacc agcctatgta cgtgttccgc aagaccgagc tgaagcactc
     3061 caagacagaa ctgaacttca aagagtggca gaaagccttc accgacgtga tgggcatgga
     3121 cgagctgtac aagggctctg ggcagtgtac taattatgct ctcttgaaat tggctggaga
     3181 tgttgagagc aacccaggtc ccggctctgg aatggcgtcc aatttcactc agtttgtgct
     3241 ggttgacaac ggcgggaccg gggacgttac ggtagccccc tcaaactttg ccaacggtat
     3301 agcggagtgg ataagcagca attctaggag tcaagcatac aaagttacat gcagcgtgcg
     3361 ccaatctagc gctcagaatc gcaagtacac cattaaagta gaggtcccca agggagcctg
     3421 gagaagctat cttaacatgg agttgaccat accaatcttc gctaccaact ctgactgtga
     3481 actcattgtg aaagccatgc aaggtctgct caaggatggt aacccaattc cgtccgctat
     3541 cgctgccaac tctgggattt acgggggcag tgggagcggt gcaggatctg gtagtccagc
     3601 tgggggagga gcaccgggta gcggtggggg gtctcagctg cacctgcccc aggttctcgc
     3661 agacgccgta tcccgccttg tactgggcaa gtttggtgat cttactgaca atttttcatc
     3721 tcctcatgcg aggcggaaag tactcgcagg cgtcgtcatg acgaccggaa ctgacgtgaa
     3781 agacgccaaa gtcatctctg tctccacggg cacaaagtgc ataaacgggg agtacatgag
     3841 cgaccggggg ctggcactga atgattgtca cgctgaaata atatctaggc gatctctgct
     3901 tagatttctc tacactcaac tcgaattgta ccttaacaac aaagatgacc agaaacgcag
     3961 tatatttcag aaatcagaac gcggcggatt tcgacttaag gaaaacgttc agttccactt
     4021 gtatatcagc acatcccctt gcggtgacgc ccgaatcttt tccccgcacg agccgatatt
     4081 ggaggagccc gcggacagac atcctaatag gaaggctaga ggccaacttc ggacgaagat
     4141 tgaaagtggc cagggtacta tcccggtgcg gtccaacgct agtattcaaa cgtgggacgg
     4201 agtccttcaa ggtgaacggc tgttgacaat gagctgctca gacaaaatcg cgcgctggaa
     4261 tgtagtggga atccaaggca gcctcttgag catattcgta gaacccatat atttctcatc
     4321 cattattttg ggctctctgt atcatggtga ccatctgtca agggctatgt accaacgaat
     4381 ttctaatatc gaggatcttc ctccactcta tacactcaat aagcctctct tgtccgggat
     4441 atcaaacgct gaggcccgcc agccagggaa agctcctaac ttcagtgtta actggaccgt
     4501 tggtgattct gcgatagagg tcatcaacgc cacgacaggt aaggatgagc tcggtagagc
     4561 ctcacgcctg tgtaaacacg cgttgtattg tagatggatg agagtacatg ggaaggtccc
     4621 atctcacttg ctccgaagca agatcactaa gcctaatgtg tatcatgagt caaaactcgc
     4681 ggctaaagaa taccaggcag ccaaagctcg actttttaca gcttttatta aggcagggct
     4741 cggggcatgg gtcgagaagc cgaccgagca ggaccaattc tctctgacgg ggagcggatc
     4801 cggctctctg ccaccattgg aaaggcttac gctgtaactt ccactggatt gtacaattac
     4861 caactgatct agaaccggtc atggccgcaa taaaatatct ttattttcat tacatctgtg
     4921 tgttggtttt ttgtgtgttc gaactagatg ctgtcgaccg atgcgctggt ctaataccca
     4981 atctctcgtc ttatccagat gttttatacg ccttaattaa tagggataac agggtaatat
     5041 cttgccctgc acgaatacca gcgacccctt gcccaaatac ttgccgtggg cctcggcctg
     5101 agagccaaaa cacttgatgc ggaagaagtc ggtgcgctcc tgcttgtcgc cggcatcgtt
     5161 gcgccacatc taggatctgc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5221 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5281 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5341 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5401 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5461 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5521 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5581 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5641 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5701 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5761 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5821 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5881 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5941 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6001 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6061 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaacca
     6121 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6181 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6241 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6301 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6361 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6421 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6481 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6541 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6601 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6661 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6721 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6781 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6841 ctttttcaat attattgaag catttatcaa ggttattgtc tcatgagcgg atacatattt
     6901 gaatgtattt agaaaaataa acaaataggg gttccgcgca cgaattggcc agcgctgcat
     6961 taccctgtta tccctaaccg g
//
